高等学校化学学报

高等学校化学学报 ›› 2000, Vol. 21 ›› Issue (8): 1187.

• 论文 • 上一篇    下一篇

溴化乙锭标记DNA电化学探针的研究

徐春, 何品刚, 方禹之   

  1. 华东师范大学化学系, 上海 200062
  • 收稿日期:1999-09-28 出版日期:2000-08-24 发布日期:2000-08-24
  • 通讯作者: 方禹之(1931年出生),男,教授,博士生导师,主要从事电分析化学研究.
  • 基金资助:

    国家自然科学基金(批准号:29575197)

Studies on the Preparation and Characterization of Ethidium Bromide Labeled DNA Probe

XU Chun, HE Pin-Gang, FANG Yu-Zhi   

  1. Department of Chemistry, East China Normal University, Shanghai 200062, China
  • Received:1999-09-28 Online:2000-08-24 Published:2000-08-24

PDF (PC)

被引次数

131

摘要： 以乙基-(3-二甲基丙基)碳化二亚胺盐酸盐(EDC)为偶联活化剂,将电化学活性物质溴化乙锭(Ethidiumbromide,EB)成功地标记在人工合成的含有21个碱基的寡聚DNA片段上,制备成EB标记DNA探针;用电化学方法将待测样品DNA片段固定在石墨电极表面,在一定的温度、pH值和离子强度条件下与EB标记DNA探针进行杂交反应,从而对靶序列DNA片段进行识别和测定.此外,还讨论了该探针的电化学性质、荧光光谱、待测DNA片段在石墨电极表面的电化学固定、DNA链碱基长度对EB标记DNA电化学探针的影响以及探针的选择性、重现性和寿命,结果令人满意.

关键词: 溴化乙锭, 电化学标记, DNA探针, 杂交反应

Abstract: The preparation and characterization of an ethidium bromide labeled DNAprobe is studied. Ethidium bromide was labeled onto a 21-bases 5'-oligonucleotides by EDC. The electrochemical characterization and fluorescence of the probe is discussed. The accumulation of sample DNAis also discussed. Besides it, the effect of base amounts on labeling and hybridization were discussed. We think the bases between 15—200 are better for electrochemical labeled DNAprobe. The EBlabeled DNAprobe was successfully used to detect sequence specific DNA. It shows a high sensitivity and selectivity. It is hoped for commodity use.

Key words: Ethidium bromide, Electrochemical label, DNA probe, Hybridization

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